US10533055B2ActiveUtilityA1

Antibodies and chimeric antigen receptors specific for CD19

Assignee: JUNO THERAPEUTICS INCPriority: Aug 28, 2014Filed: Aug 28, 2015Granted: Jan 14, 2020

Est. expiryAug 28, 2034(~8.1 yrs left)· nominal 20-yr term from priority

Inventors:Yan Chen Steven M. Shamah Csaba Pazmany Jui Dutta-Simmons

A61P 35/02A61P 35/00C07K 14/7051C07K 16/2803C07K 2317/94C07K 2317/622C07K 2317/92C07K 2317/34C07K 2317/21C07K 2317/73C07K 2319/33C07K 14/70521C07K 2319/00C07K 2317/56C07K 2317/565C07K 2317/53C07K 16/2896A61K 2039/505A61K 40/11A61K 40/31A61K 40/4211A61K 2239/38A61K 2239/31A61K 2239/48C07K 2319/03C12N 5/0636C07K 2319/02A61K 39/395

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Claims

Abstract

Provided are CD19 binding molecules, including anti-CD19 antibodies, including antibody fragments such as single-chain fragments, and chimeric receptors including the antibodies, such as chimeric antigen receptors (CARs). Among the antibodies are human antibodies, including those that compete for binding to CD19 with reference antibodies, such as murine antibodies. In some embodiments, the antibodies display similar functional properties to the reference antibodies, such as comparable binding affinities and/or competitive inhibition properties. Also provided are genetically engineered cells expressing the chimeric receptors, and uses of the binding molecules and cells adoptive cell therapy.

Claims

exact text as granted — not AI-modified

What is claimed:

1. A chimeric antigen receptor (CAR) comprising:
a) an extracellular portion comprising an anti-CD19 antibody or antigen-binding fragment thereof, wherein said antibody or antigen-binding fragment comprises:
a heavy chain variable (VH) region, wherein the VH region comprises a heavy chain complementarity determining region 1 (CDR-H1) comprising the amino acid sequence set forth in SEQ ID NO: 18; a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 81; and a CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 20; and
a light chain variable (VL) region, wherein the VL region comprises:
a light chain complementarity determining region 1 (CDR-L1) comprising the amino acid sequence set forth in SEQ ID NO: 21, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 22, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 23; or
a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 21, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 22, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 24; or
a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 25, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 26, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 27; or
a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 28, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 29, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 30; or
a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 31, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 32, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 33;
and

b) an intracellular signaling domain, wherein the intracellular signaling domain comprises an immunoreceptor tyrosine-based activation motif (ITAM).

2. The chimeric antigen receptor of claim 1 , wherein the antibody or antigen binding fragment thereof comprises:
a VH region comprising the amino acid sequence set forth in SEQ ID NO: 11, or a sequence that has at least 90% amino acid sequence identity to SEQ ID NO: 11; and a VL region comprising the amino acid sequence set forth in SEQ ID NO: 13;
a VH region comprising the amino acid sequence set forth in SEQ ID NO: 11, or a sequence having at least 90% amino acid sequence identity to SEQ ID NO:11, and a VL region comprising the amino acid sequence set forth in SEQ ID NO: 14, or a sequence having at least 90% amino acid sequence identity to SEQ ID NO:14;
a VH region comprising the amino acid sequence set forth in SEQ ID NO: 11, or a sequence having at least 90% amino acid sequence identity to SEQ ID NO:11, and a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16, or a sequence having at least 90% amino acid sequence identity to SEQ ID NO:16;
a VH region comprising the amino acid sequence set forth in SEQ ID NO: 12, or a sequence having at least 90% amino acid sequence identity to SEQ ID NO:12, and a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17, or a sequence having at least 90% amino acid sequence identity to SEQ ID NO:17; or
a VH region comprising the amino acid sequence set forth in SEQ ID NO: 12, or a sequence having at least 90% amino acid sequence identity to SEQ ID NO:12, and a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15, or a sequence having at least 90% amino acid sequence identity to SEQ ID NO:15.

3. A chimeric antigen receptor (CAR), comprising:
a) an extracellular portion comprising an anti-CD19 antibody or antigen-binding fragment thereof, said antibody or antigen-binding fragment comprising:
a heavy chain variable (VH) region comprising a CDR-H1, a CDR-H2, and a CDR-H3, contained within the VH region amino acid sequence set forth in SEQ ID NO: 11; and
a light chain variable (VL) region comprising a CDR-L1, a CDR-L2, and a CDR-L3, contained within the VL region amino acid sequence set forth in SEQ ID NO: 13, 14, 15, 16 or 17 and
b) an intracellular signaling domain, wherein the intracellular signaling domain comprises an immunoreceptor tyrosine-based activation motif (ITAM).

4. The chimeric antigen receptor of claim 1 , wherein the antibody or antigen binding fragment thereof comprises a VH region comprising the amino acid sequence set forth in SEQ ID NO: 11, or a sequence having at least 90% amino acid sequence identity to SEQ ID NO:11, and a VL region comprising the amino acid sequence set forth in SEQ ID NO: 13, or a sequence having at least 90% amino acid sequence identity to SEQ ID NO:13.

5. The chimeric antigen receptor of claim 1 , wherein the antibody or antigen binding fragment thereof comprises a VH region comprising the amino acid sequence set forth in SEQ ID NO: 11, or a sequence having at least 90% amino acid sequence identity to SEQ ID NO:11, and a VL region comprising the amino acid sequence set forth in SEQ ID NO: 14, or a sequence having at least 90% amino acid sequence identity to SEQ ID NO:14.

6. The chimeric antigen receptor of claim 1 , wherein the antibody or antigen binding fragment thereof comprises a VH region comprising the amino acid sequence set forth in SEQ ID NO: 11, or a sequence having at least 90% amino acid sequence identity to SEQ ID NO:11, and a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16, or a sequence having at least 90% amino acid sequence identity to SEQ ID NO:16.

7. The chimeric antigen receptor of claim 1 , wherein:
the VH and VL regions of the antibody or fragment comprise the amino acid sequences set forth in SEQ ID NOs: 12 and 17, respectively;
the VH and VL regions of the antibody or fragment comprise the amino acid sequences set forth in SEQ ID NOs: 12 and 15, respectively;
the VH and VL regions of the antibody or fragment comprise the amino acid sequences set forth in SEQ ID NOs: 11 and 13, respectively;
the VH and VL regions of the antibody or fragment comprise the amino acid sequences set forth in SEQ ID NOs: 11 and 14, respectively; or
the VH and VL regions of the antibody or fragment comprise the amino acid sequences set forth in SEQ ID NOs: 11 and 16, respectively.

8. The chimeric antigen receptor of claim 1 , wherein the anti-CD19 antibody or antigen-binding fragment specifically binds to human CD19.

9. The chimeric antigen receptor of claim 1 , wherein the antibody or antigen-binding fragment is human.

10. The chimeric antigen receptor of claim 1 , wherein the antibody or antigen-binding fragment is a single chain molecule.

11. The chimeric antigen receptor of claim 10 , wherein the single chain molecule comprises the antibody variable regions joined by a flexible linker.

12. The chimeric antigen receptor of claim 10 , wherein the single chain molecule comprises an scFv.

13. The chimeric antigen receptor of claim 12 , wherein the scFv comprises a linker comprising the sequence set forth SEQ ID NO: 34.

14. The chimeric antigen receptor of claim 12 , wherein the scFv comprises a sequence that exhibits at least 95% sequence identity to the amino acid sequence set forth in any one of SEQ ID NOS: 2, 4, 6, 8, and 10.

15. The chimeric antigen receptor of claim 12 , wherein the scFv comprises the amino acid sequence set forth in any one of SEQ ID NOS: 2, 4, 6, 8 and 10.

16. The chimeric antigen receptor of claim 1 , wherein the intracellular signaling domain comprises a signaling domain of a zeta chain of a CD3-zeta (CD3ζ) chain.

17. The chimeric antigen receptor of claim 16 , wherein the intracellular signaling domain further comprises an intracellular signaling region of a T cell costimulatory molecule.

18. The chimeric antigen receptor of claim 3 , wherein the intracellular signaling domain comprises a signaling domain of a zeta chain of a CD3-zeta (CD3ζ) chain.

19. The chimeric antigen receptor of claim 18 , wherein the intracellular signaling domain further comprises an intracellular signaling region of a T cell costimulatory molecule.

20. A conjugate, comprising:
a) an anti-CD19 antibody or antigen-binding fragment thereof, wherein said antibody or antigen-binding fragment comprises:
a heavy chain variable (VH) region, wherein the VH region comprises a heavy chain complementarity determining region 1 (CDR-H1) comprising the amino acid sequence set forth in SEQ ID NO: 18; a CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 81; and a CDR-H3 comprising the amino acid sequence set forth as SEQ ID NO: 20; and
and a light chain variable (VL) region, wherein the VL region comprises:
a light chain complementarity determining region 1 (CDR-L1) comprising the amino acid sequence set forth in SEQ ID NO: 21, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 22, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 23; or
a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 21, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 22, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 24; or
a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 25, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 26, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 27; or
a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 28, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 29, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 30; or
a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 31, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 32, and a CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 33; and

b) and a heterologous molecule or moiety.

21. The conjugate of claim 20 , wherein the conjugate is a chimeric antigen receptor (CAR).

22. The chimeric antigen receptor of claim 1 , wherein the CDR-H2 comprises the amino acid sequence set forth in SEQ ID NO: 19 (GISWNSGRIGYADSVKG).

23. The chimeric antigen receptor of claim 3 , wherein the anti-CD19 antibody or antigen-binding fragment specifically binds to human CD19.

24. The chimeric antigen receptor of claim 1 , wherein the intracellular signaling domain further comprises an intracellular signaling region of a T cell costimulatory molecule.

25. The chimeric antigen receptor of claim 3 , wherein the intracellular signaling domain further comprises an intracellular signaling region of a T cell costimulatory molecule.

26. The conjugate of claim 20 , wherein the CDR-H2 comprises the amino acid sequence set forth in SEQ ID NO: 19 (GISWNSGRIGYADSVKG).

27. The chimeric antigen receptor of claim 1 , wherein the antibody or antigen binding fragment thereof comprises a VH region comprising the amino acid sequence set forth in SEQ ID NO: 12, or a sequence having at least 90% amino acid sequence identity to SEQ ID NO:12, and a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17, or a sequence having at least 90% amino acid sequence identity to SEQ ID NO:17.

28. The chimeric antigen receptor of claim 1 , wherein the antibody or antigen binding fragment thereof comprises a VH region comprising the amino acid sequence set forth in SEQ ID NO: 12, or a sequence having at least 90% amino acid sequence identity to SEQ ID NO:12, and a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15, or a sequence having at least 90% amino acid sequence identity to SEQ ID NO:15.

29. The chimeric antigen receptor of claim 16 , wherein the antibody or antigen binding fragment thereof comprises:
a VH region comprising the amino acid sequence set forth in SEQ ID NO: 11, or a sequence having at least 90% amino acid sequence identity to SEQ ID NO:11, and a VL region comprising the amino acid sequence set forth in SEQ ID NO: 13, or a sequence having at least 90% amino acid sequence identity to SEQ ID NO:13;
a VH region comprising the amino acid sequence set forth in SEQ ID NO: 11, or a sequence having at least 90% amino acid sequence identity to SEQ ID NO:11, and a VL region comprising the amino acid sequence set forth in SEQ ID NO: 14, or a sequence having at least 90% amino acid sequence identity to SEQ ID NO:14;
a VH region comprising the amino acid sequence set forth in SEQ ID NO: 11, or a sequence having at least 90% amino acid sequence identity to SEQ ID NO:11, and a VL region comprising the amino acid sequence set forth in SEQ ID NO: 16, or a sequence having at least 90% amino acid sequence identity to SEQ ID NO:16;
a VH region comprising the amino acid sequence set forth in SEQ ID NO: 12, or a sequence having at least 90% amino acid sequence identity to SEQ ID NO:12, and a VL region comprising the amino acid sequence set forth in SEQ ID NO: 17, or a sequence having at least 90% amino acid sequence identity to SEQ ID NO:17; or
a VH region comprising the amino acid sequence set forth in SEQ ID NO: 12, or a sequence having at least 90% amino acid sequence identity to SEQ ID NO:12, and a VL region comprising the amino acid sequence set forth in SEQ ID NO: 15, or a sequence having at least 90% amino acid sequence identity to SEQ ID NO:15.

30. The chimeric antigen receptor of claim 29 , wherein the intracellular signaling domain further comprises an intracellular signaling region of a T cell costimulatory molecule.

31. The chimeric antigen receptor of claim 29 , wherein:
the VH and VL regions of the antibody or fragment comprise the amino acid sequences set forth in SEQ ID NOs: 11 and 13, respectively;
the VH and VL regions of the antibody or fragment comprise the amino acid sequences set forth in SEQ ID NOs: 11 and 14, respectively;
the VH and VL regions of the antibody or fragment comprise the amino acid sequences set forth in SEQ ID NOs: 11 and 16, respectively;
the VH and VL regions of the antibody or fragment comprise the amino acid sequences set forth in SEQ ID NOs: 12 and 15, respectively; or
the VH and VL regions of the antibody or fragment comprise the amino acid sequences set forth in SEQ ID NOs: 12 and 17, respectively.

32. The chimeric antigen receptor of claim 29 , wherein the antibody or antigen binding fragment comprises an scFv.

33. The chimeric antigen receptor of claim 32 , wherein the scFv comprises a sequence that exhibits at least 95% sequence identity to the amino acid sequence set forth in any one of SEQ ID NOS: 2, 4, 6, 8, or 10.

34. The chimeric antigen receptor of claim 1 , wherein the VH region comprises a CDRH1, a CDRH2 and a CDRH3 set forth in SEQ ID NOS: 18, 19 and 20, respectively and the VL region comprises a CDRL1, a CDRL2 and a CDRL3 set forth in SEQ ID NOS: 21, 22 and 23, respectively.

35. The chimeric antigen receptor of claim 34 , wherein the intracellular signaling domain comprises a signaling domain of a zeta chain of a CD3-zeta (CD3ζ) chain.

36. The chimeric antigen receptor of claim 35 , wherein the intracellular signaling domain further comprises an intracellular signaling region of a T cell costimulatory molecule.

37. The chimeric antigen receptor of claim 1 , wherein the VH region comprises a CDRH1, a CDRH2 and a CDRH3 set forth in SEQ ID NOS: 18, 19 and 20, respectively and the VL region comprises a CDRL1, a CDRL2 and a CDRL3 set forth in SEQ ID NOS: 21, 22 and 24, respectively.

38. The chimeric antigen receptor of claim 37 , wherein the intracellular signaling domain comprises a signaling domain of a zeta chain of a CD3-zeta (CD3ζ) chain.

39. The chimeric antigen receptor of claim 38 , wherein the intracellular signaling domain further comprises an intracellular signaling region of a T cell costimulatory molecule.

40. The chimeric antigen receptor of claim 1 , wherein the VH region comprises a CDRH1, a CDRH2 and a CDRH3 set forth in SEQ ID NOS: 18, 19 and 20, respectively and the VL region comprises a CDRL1, a CDRL2 and a CDRL3 set forth in SEQ ID NOS: 25, 26 and 27, respectively.

41. The chimeric antigen receptor of claim 40 , wherein the intracellular signaling domain comprises a signaling domain of a zeta chain of a CD3-zeta (CD3ζ) chain.

42. The chimeric antigen receptor of claim 41 , wherein the intracellular signaling domain further comprises an intracellular signaling region of a T cell costimulatory molecule.

43. The chimeric antigen receptor of claim 1 , wherein the VH region comprises a CDRH1, a CDRH2 and a CDRH3 set forth in SEQ ID NOS: 18, 19 and 20, respectively and the VL region comprises a CDRL1, a CDRL2 and a CDRL3 set forth in SEQ ID NOS: 28, 29 and 30, respectively.

44. The chimeric antigen receptor of claim 43 , wherein the intracellular signaling domain comprises a signaling domain of a zeta chain of a CD3-zeta (CD3ζ) chain.

45. The chimeric antigen receptor of claim 44 , wherein the intracellular signaling domain further comprises an intracellular signaling region of a T cell costimulatory molecule.

46. The chimeric antigen receptor of claim 1 , wherein the VH region comprises a CDRH1, a CDRH2 and a CDRH3 set forth in SEQ ID NOS: 18, 19 and 20, respectively and the VL region comprises a CDRL1, a CDRL2 and a CDRL3 set forth in SEQ ID NOS: 31, 32 and 33, respectively.

47. The chimeric antigen receptor of claim 46 , wherein the intracellular signaling domain comprises a signaling domain of a zeta chain of a CD3-zeta (CD3ζ) chain.

48. The chimeric antigen receptor of claim 47 , wherein the intracellular signaling domain further comprises an intracellular signaling region of a T cell costimulatory molecule.

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